Received:16 July 2019,
Published:25 December 2019
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Xingxin LI, Anmao LI, Chensi ZENG, et al. Silencing NUPR1 down-regulates autophagy and promotes migration and invasion in human multiple myeloma U266 cells[J]. Journal of Third Military Medical University, 2019, 41(23): 2322-2327.
Xingxin LI, Anmao LI, Chensi ZENG, et al. Silencing NUPR1 down-regulates autophagy and promotes migration and invasion in human multiple myeloma U266 cells[J]. Journal of Third Military Medical University, 2019, 41(23): 2322-2327. DOI: 10.16016/j.1000-5404.201907123.
Objective
2
To explore the effect of silencing nuclear protein 1 (NUPR1) on autophagy
migration and invasion in human multiple myeloma (MM) U266 cells and its possible mechanism.
Methods
2
NUPR1-shRNA was transfected into MM U266 cells
and the transfection rate was detected by flow cytometry. Then the cells were divided into negative control cells (transfected with unrelated sequence RNA lentiviral vector) and NUPR1 knockdown cells (transfected with NUPR1-shRNA lentiviral vector). The interference effect of NUPR1 was detected by qRT-PCR
and the expression levels of autophagy related proteins (Beclin1
ATG5
LC3Ⅱ/LC3Ⅰ
P62)
migration and invasion associated proteins (MMP9
CXCR4) and pathway proteins (p-AKT/T-AKT
p-mTOR/T-mTOR) were measured by Western blotting. Transmission electron microscopy (TEM) was used to observe autophagosomes. Transwell test was employed to detect cell migration and invasion.
Results
2
The expression of NUPR1 at mRNA and protein level was significantly decreased in NUPR1 knockdown cells (
P
<
0.05). The protein levels of autophagy related proteins
ATG5
Beclin1 and LC3Ⅱ/LC3Ⅰwere significantly lower
while those of f MMP9
CXCR4
P62
p-AKT/T-AKT and p-mTOR/T-mTOR were increased obviously in the NUPR1 knockdown cells than the negative control cells (
P
<
0.05). There were significantly reduced autophagosomes in NUPR1 knockdo
wn cells. While
migratory and invasive abilities were enhanced in NUPR1 knock down group
and rapamycin reduced the promoting effects.
Conclusion
2
Silencing NUPR1 may promotes migration and invasion by down-regulating autophagy in U266 cells
which may be related to activation of the AKT/mTOR pathway.
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